ProjectSummary/Abstract Thisprojectwilldevelopanautomatedcapillaryelectrophoresis-tandemmassspectrometryplatformforthe structuralanalysisofmixturesofglycosaminoglycanoligomersthatareisolatedinproteinbindingassays.This classofcarbohydratescanbefoundonthesurfaceofallanimalcells,andplayakeyroleinmanyimportant biologicalprocesses.Glycosaminoglycansarelinear,polysulfatedcopolymers,typicallywitharepeating disaccharideunitcomposedofanN-acetyl-hexosamineandglucuronicacid,andcanbemorethan200repeat unitslong.Modificationscanoccuranywherealongthechain,andtheseincludeO-sulfation,N- deacetylation/N-sulfation,andepimerizationoftheuronicacid.Theseareaparticularlychallengingclassof compoundstocharacterizeduetotheirheterogeneousstructures,anoutcomeoftheirnon-template biosynthesis.Nevertheless,withinthelongglycanchainsareshortregionsofdefinedmodificationthattarget specificproteinswithremarkablespecificity.Theseglycan-proteininteractionsarethebasisforthebiological activityoftheglycosaminoglycan.Thereisacriticalneedfortoolstofindtheseorganizedregionswiththelong glycanchain,andtodeterminetheirpatternofmodification.Afewspecializedlaboratorieshavedeveloped massspectrometrymethodsforthestructuralanalysisofglycosaminoglycans,butthesehavenotfoundtheir wayintobiologicallaboratoriesatlarge.Thegoalofthisprojectistoproduceaplatformwithahighdegreeof automationsothatitcanbedeployedinthelaboratoriesofbiologyresearchers.Theaccessibilityofthese advancedmassspectrometrymethodswillresultfrominnovationsinhardware,software,andmethods development.